TY  - JOUR
A1  - Fischer, Klaus
A1  - Wacht, Marion
T1  - Fluorimetric microplate assay for the determination of extracellular alkaline phosphatase kinetics and inhibition kinetics in activated sludge
T2  - MethodsX
N2  - The microbial enzyme alkaline phosphatase contributes to the removal of organic phosphorus compounds from wastewaters. To cope with regulatory threshold values for permitted maximum phosphor concentrations in treated wastewaters, a high activity of this enzyme in the biological treatment stage, e.g., the activated sludge process, is required. To investigate the reaction dynamics of this enzyme, to analyze substrate selectivities, and to identify potential inhibitors, the determination of enzyme kinetics is necessary. A method based on the application of the synthetic fluorogenic substrate 4-methylumbelliferyl phosphate is proven for soils, but not for activated sludges. Here, we adapt this procedure to the latter. The adapted method offers the additional benefit to determine inhibition kinetics. In contrast to conventional photometric assays, no particle removal, e.g., of sludge pellets, is required enabling the analysis of the whole sludge suspension as well as of specific sludge fractions. The high sensitivity of fluorescence detection allows the selection of a wide substrate concentration range for sound modeling of kinetic functions.
- Fluorescence array technique for fast and sensitive analysis of high sample numbers
- No need for particle separation – analysis of the whole (diluted) sludge suspension
- Simultaneous determination of standard and inhibition kinetics
KW  - Enzyme kinetics
KW  - Enzyme inhibition
KW  - Enzymatic reactions
KW  - Biological wastewater treatment
KW  - Organic phosphorus compounds
KW  - Phosphor elimination
KW  - Belebtschlamm
KW  - Abwasserreinigung
KW  - Phosphororganische Verbindungen
KW  - Phosphatelimination
KW  - Enzymkinetik
KW  - Enzymimmunassay
Y1  - 2023
UR  - https://ubt.opus.hbz-nrw.de/frontdoor/index/index/docId/2109
UR  - https://nbn-resolving.org/urn:nbn:de:hbz:385-1-21093
VL  - 2023
IS  - Band 11
PB  - Elsevier
CY  - Amsterdam
ER  -